This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. CORE B: Cell and Molecular Biology CORE CORE B shall meet all of the cell culture, molecular biology, and fluorescence-activated cell sorting (FACS) needs of the COBRE PIs who are not routinely using these methods in their own laboratories. CORE B can provide stem and/or lineage restricted precursor cells derived from embryonic cortex or spinal cord. These can be from fetuses that constitutively express endogenous labels such as green fluorescent protein (GFP) or human placental alkaline phosphatase (hPLP) or they can be exogenous labeled or induced to express transgenes as described below. CORE B can provide retroviral, adenoviral, and lentiviral vectors that express whatever cDNA the Investigators'need. In addition, we have regulatable retroviruses (TetON and TetOFF) that are responsive to doxycyline, both in vitro and in vivo. CORE B has developed lentiviruses that express siRNA cassettes that we can target any mRNA that the COBRE Investigators'want silenced. CORE B has also developed RNAse protection assays that can be modified to all Investigators'needs. CORE B can provide expertise in quantiative real time PCR to assess mRNA levels for any gene of interest. All of these reagents have been characterized by the CORE prior to their dispersal to the COBRE PIs to ensure quality control.